Coexisting cancer stem cells with heterogeneous gene amplifications, transcriptional profiles, and malignancy are isolated from single glioblastomas

Glioblastoma (GBM) is known as an intractable, highly heterogeneous tumor encompassing multiple subclones, each supported by a distinct glioblastoma stem cell (GSC). The contribution of GSC genetic and transcriptional heterogeneity to tumor subclonal properties is debated. In this study, we describe the systematic derivation, propagation, and characterization of multiple distinct GSCs from single, treatment-naive GBMs (GSC families). The tumorigenic potential of each GSC better correlates with its transcriptional profile than its genetic make-up, with classical GSCs being inherently more aggressive and mesenchymal more dependent on exogenous growth factors across multiple GBMs. These GSCs can segregate and recapitulate different histopathological aspects of the same GBM, as shown in a paradigmatic tumor with two histopathologically distinct components, including a conventional GBM and a more aggressive primitive neuronal component. This study provides a resource for investigating how GSCs with distinct genetic and/or phenotypic features contribute to individual GBM heterogeneity and malignant escalation. [Display omitted] •Distinct GSCs can be derived from single GBMs by different growth factor cocktails•GSCs segregate different RTK expression and gene amplifications•GSCs segregate distinct transcriptional profiles, each related to different malignancy•GSCs with a neural progenitor profile recapitulate the GBM primitive neuronal component De Bacco et al. devise a methodology to propagate distinct glioblastoma stem-like cells (GSCs) from individual tumors, stably segregating different features coexisting in the original tumor. The GSCs' transcriptional profile is a superior indicator of their tumorigenicity compared with their genetic make-up, uncovering diverse phenotypes that drive intratumoral heterogeneity.